Nano-enzyme and preparation method thereof and application thereof in determination of 5-hydroxytryptamine
A technology of hydroxytryptamine and nanozyme, which is applied in chemical instruments and methods, measuring devices, fluorescence/phosphorescence, etc., and can solve problems such as high price, expensive chromatography equipment, and long analysis cycle
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Embodiment 1
[0026] Example 1 Preparation of nanozyme RF Cu Tb DDE
[0027] Dissolve 1.5 mg riboflavin (RF) in 10 mL water and heat to 80°C to dissolve, add 1 mL 10 mM Tb(NO 3 ) 3 aqueous solution and 1 mL of 10 mM Cu(NO 3 ) 2 Add 4 mL of ethylene glycol solution containing 0.04 g of polyvinylpyrrolidone (PVP, molecular weight: 40,000), and add 4 mL of 5 mM 4,4-dicarboxydiphenyl ether DMF solution to form terbium ion: copper ion: nucleus The molar ratio of flavin: 4, 4-dicarboxydiphenyl ether is a mixture of 1:1:0.4:2. After stirring vigorously for 20 minutes, the mixture was transferred to the reaction kettle, reacted at 160 °C for 6 hours, cooled to room temperature (25 °C), collected the precipitate by centrifugation, washed with ethanol for 3 times, and dried in an oven at 80 °C to obtain nanozyme. figure 1 It is a scanning electron micrograph of the prepared nanozyme RF Cu Tb DDE, and the nanozyme RF Cu Tb DDE is a hollow spherical nanoparticle with a particle size of 50-300 nm. ...
Embodiment 2
[0028] Example 2 Nanozyme RF Cu Tb DDE Catalyzes 5-Hydroxytryptamine
[0029] Take 20 μL of 0.02 mM 5-hydroxytryptamine aqueous solution and 10 μL of the nanozyme RF Cu TbDDE suspension (0.2 mg / mL) prepared in Example 1, and add them to 970 μL of water samples respectively. UV-Vis and fluorescence spectra of the time-determined solutions. figure 2 It is the ultraviolet-visible spectrum of the solution at 0, 5, 10, 15, 20, 25, 30, 35, and 40 minutes of reaction. Under the catalysis of nanozyme RF Cu Tb DDE, the solution is at 220 nm, 276 nm, and 297 nm. The absorbance increased continuously with the reaction, and the peaks at 220 nm, 276 nm and 297 nm were the absorption peaks of 5-hydroxyindoleacetic acid, indicating the formation of 5-hydroxyindoleacetic acid. image 3 It is the fluorescence spectrogram of the solution at 0, 5, 10, 15, 20, 25, 30, 35, and 40 minutes. Under the catalysis of nanozyme RF Cu Tb DDE, the fluorescence intensity of the solution at 336 nm increases...
Embodiment 3
[0030] Example 3 Nanozyme RF Cu Tb DDE Luminescent Detection of 5-Hydroxytryptamine
[0031]Take 10 μL of the nanozyme RF Cu Tb DDE (0.3 mg / mL) prepared in Example 1 and add it to 980 μL of HEPES buffer solution (10 mM, pH 7). Add 10 μL of different concentrations of 5-hydroxytryptamine aqueous solution (containing 10% ethanol, v / v) to the above buffer to prepare 5-hydroxytryptamine containing 0, 0.05, 0.1, 0.2, 0.5 and 1.0 μM Standard solution, mix well and react for 30 minutes, measure Tb under 278 nm excitation 3+ Fluorescence intensity at 545 nm. Figure 5 is Tb 3+ Plot of fluorescence intensity versus 5-hydroxytryptamine concentration, Tb 3+ The fluorescence intensity is linearly proportional to the concentration of 5-hydroxytryptamine, and the detection sensitivity is 0.05 μM. The content of 5-hydroxytryptamine in the sample solution is obtained by using the linear relationship diagram and the measured fluorescence intensity of the 5-hydroxytryptamine sample solutio...
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